| Products/Services Used | Details | Operation |
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| Recombinant Proteins | In the present study, human miR-10-5p was transfected into lentivirus (lentimiR10a-5p) and empty lentivirus lenti-miRNA control (mi-RC) was used as a negative control. All products were purchased from SunBio Biotech, Co., Ltd. (Beijing, China). Lenti-miR-10a-5p or lenti-miRC [100 nM; GenScript (Nanjing) Co., Ltd., Nanjing, China] were then transduced into HeLa and SiHa cells using Lipofectamine 2000 (Biomics Biotechnologies Co., Ltd., Nantong, China) following a previously published protocol (18).The concentrations of stacking and resolving gel were 5 and 15% respectively. Following SDS-PAGE, proteins were transferred to PVDF membranes [cat no. L03014; GenScript (Nanjing) Co., Ltd.]. Blocking was performed with 5% dry milk for 1 h at room temperature. Rabbit anti-BDNF antibody (1:200, cat no. sc-20981; Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) was used as the primary antibody and was incubated at 4˚C overnight. Goat anti‑rabbit horseradish‑peroxidase conjugated antibody [1:1,000 dilution, cat no. A00098; GenScript (Nanjing) Co., Ltd.] was used as the secondary antibody. | Get A Quote |
The aim of the present study was to investigate the effect and mechanism of microRNA (miR)-10a-5p in human cervical cancer. The expression level of miR-10-5p in cervical cancer lines was assessed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). In cervical cancer HeLa and SiHa cells， miR-10-5p was ectopically overexpressed by lentiviral transduction. Brain-derived neurotrophic factor (BDNF) was then overexpressed in HeLa and SiHa cells to evaluate its selective effect on miR-10-5p in cervical cancer modulation. The targeting of miR-10-5p on its downstream gene， BDNF， was evaluated using RT-qPCR and western blot analysis. Cervical cancer cell viability and cell cycle was evalua... More
